Imperial College/22 July 2008

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22 July 2008

Debriefing

Each Group gave a summary of their sections, followed by a discussion on any potential problems:

B.subtilis

  • The group has found an amazing database of promoters and encoding regions of the B.subtilis genome. Several promoters have been investigated, the group was unclear if new terminators other than the E.coli terminators will be required.

Light Induction

  • The natural light detecting pathway has been investigated and the relevant parts required have been investigated. In addition, parameters for the modelling have been investigated including response time.
  • The only remaining issue is to try to engineer a light detecting pathway in B.suntilis.

Biomaterials

  • Secretion paths have been investigated by the biomaterials group. The main transport system used in B.suntilis is the SRP transport system. The signal peptide used is a variable sequence, however several signal peptides are commonly used. The group has suggested using 3 signal peptides in parallel with the 3 proposed biomaterials leading to 9 potential constructs, however after discussion the group agreed it would be better to use just one signal peptide for each biomaterial leading to just 3 potential constructs.
  • One key unresolved question is how the signal peptide is cleaved, needs to be resolved


  • Biomaterials have been chosen to pursue, these include a range of naturally occurring and synthetic peptides. The group discussed about using the mass spectrometry facilities at Imperial to confirm the presence of the biomaterial and levels of other secreted proteins.

Summary

  • Key issue is to work out a strategy for assembly of our constructs.

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